[Effects of moxibustion at "Tianshu"（ST25） and "Shangjuxu" （ST37） on colonic metabolites and inflammatory factors in rats with Crohn's disease].

OBJECTIVE: To observe the effects of moxibustion at "Tianshu"（ST25） and "Shangjuxu"（ST37） on the colonic metabolites and inflammatory factors in rats with Crohn's disease（CD）, so as to explore the mechanisms of moxibustion in protecting colon of CD rats based on metabolomics. METHODS: Twelve rats were first randomly selected from 36 male SD rats as a normal group（NG）. The CD model was induced by 2, 4, 6 trinitrobenzene sulfonic acid（TNBS） enema on the rest 24 rats. After successful modeling, rats were randomly divided into model（TNBS） and moxibustion（TNBS+MOX） groups（n=10 rats/group）. Moxibustion was applied at bilateral ST25 and ST37 for 30 min, once daily for 7 consecutive days in the TNBS+MOX group, while rats in the NG and TNBS groups did not receive any interventions. Body weight of rats was recorded and disease activity index（DAI） was assessed during the experiment. After interventions, HE staining was performed to observe pathological damage of colon. Serum levels of inflammatory factors were measured by ELISA. NMR hydrogen spectroscopy was used to detect colonic metabolites of each group, and orthogonal partial least squares discriminant analysis（OPLS-DA） was used to screen differential colonic metabolites between groups, followed by pathway analysis using MetaboAnalyst 5.0 platform. RESULTS: After modeling, compared with the NG group, the body weight of the rats in the TNBS group was significantly decreased（P<0.05）, the DAI score was increased （P<0.05）, the colon had obvious inflammatory damage and the pathological injury index was increased（P<0.05）, and levels of serum tumor necrosis factor-α（TNF-α）, interleukin（IL）-1ß and interferon-γ（IFN-γ） were significantly increased（P<0.05）. After moxibustion intervention, compared with the TNBS group, the body weight was significantly increased（P<0.05）, while the levels of serum TNF-α, IL-1ß, IFN-γ, and DAI score of the rats in the TNBS+MOX group were significantly decreased（P<0.05）, with alleviated colonic inflammatory injury detected by HE staining. Compared with the NG group, the relative expressions of colonic hypoxanthine, betaine, creatine, inositol, taurine, uracil, and methanol of the TNBS group were decreased（P<0.05）, while the relative expressions of histidine, leucine, proline, lysine, isoleucine, phenylalanine, tyrosine, propionic acid, and valine were increased（P<0.05） in the TNBS group, among which, relative expressions of hypoxanthine, leucine, lysine, isoleucine, betaine, tyrosine, and taurine were reversed in the TNBS+MOX group relevant to the TNBS group, mainly involving phenylalanine, tyrosine and tryptophan biosynthesis, and taurine and subtaurine metabolism pathway. CONCLUSION: The mechanism of moxibustion at ST25 and ST37 for CD may be related to improving colon metabolic disorder state by regulating multiple metabolic metabolites and metabolic pathways, and reducing the level of inflammatory factors, so as to maintain intestinal immune homeostasis.

Effects of moxibustion and acupuncture at Zusanli (ST 36) and Zhongwan (CV 12) on chronic atrophic gastritis in rats.

OBJECTIVE: To evaluate the effects of moxibustion and acupuncture of Zusanli (ST 36) and Zhongwan (CV 12) acupoints on chronic atrophic gastritis (CAG) in rats, and to study the mechanisms behind their actions. METHODS: Forty-four male Sprague-Dawley rats were induced with CAG by intragastric administration of 40% ethanol combined with free drinking of N-methyl-N'nitro-N-nitrosoguanidine and irregular feeding for 12 weeks, followed by daily treatment with moxibustion or acupuncture for 2 weeks. Histopathologic examination, Western blotting of cytokines [epidermal growth factor (EGF), EGF receptor (EGFR), extracellular signal-regulated kinase (ERK), phosphorylated ERK (p-ERK)], and 1H NMR-based metabolic profiling of gastric tissues were used to measure changes related to CAG modeling and treatment. RESULTS: Moxibustion and acupuncture at Zusanli (ST 36) and Zhongwan (CV 12) each relieved CAG-induced abnormalities in histopathology and cytokine expression of ERK and p-ERK. Only moxibustion treatment regulated the expression of EGF and EGFR. Metabolites that were increased in gastric tissue by CAG induction (alanine, nicotinamide adenine dinucleotide phosphate, uracil DNA glycosylase, lactate, glycerol and adenosine) were restored to normal levels after moxibustion treatment; acupuncture treatment only normalized the levels of adenosine monophosphate and glycerol. CONCLUSION: Our findings suggest that moxibustion or acupuncture at Zusanli (ST 36) and Zhongwan (CV 12) can significantly improve the condition of CAG in rats. These treatments exert their effects on CAG through different mechanisms.

[Observation of electroacupuncture intervention on cell apoptosis and Bax and Bcl-2 expression in cerebral and myocardial tissues in cerebral ischemia rats based on "heart-brain correlation" theory].

OBJECTIVE: To compare the effect of electroacupuncture (EA) of "Neiguan" (PC6) of the Pericardium Meridian, "Shenmen" (HT7) of the Heart Meridian，"Shuigou" (GV26) of the Governor Vessel and "Zhaohai" (KI6) of the Kidney Meridian on myocardial and cerebral cell apoptosis in cerebral ischemia (CI) rats, so as to explore its mechanism underlying improvement of CI based on the theory of "Heart-brain Correlation". METHODS: Forty-eight SD rats (half male and half female) were randomly divided into normal control, model, PC6, HT7, GV26 and KI6 groups (n＝8 in each one). The CI model was established by occlusion of the middle cerebral artery (MCAO). EA (4 Hz/20 Hz, 1.5 mA) was applied to the right PC6, HT7, GV26 or KI6 respectively for 30 min, once every 12 h for 5 times. The cell apoptosis of the ischemic myocardial and cerebral tissues was detected by TUNEL method, and the expression of cerebral and myocardial Bax and Bcl-2 was determined by using immunohistochemistry. RESULTS: Following modeling, the cell apoptosis percentages and Bax-positive cells of both myocardial and cerebral tissues were significantly increased in the model group in comparison with the control group (P<0.01). After EA intervention, the cerebral apoptotic percentage and cerebral Bax-positive cells in the cerebral tissue of the PC6, HT7 and GV26 groups, and the myocardial apoptosis percentage in the PC6 and HT7 groups, as well as the myocardial Bax-positive cells in the PC6 group were obviously decreased (P<0.05, P<0.01), while the cerebral Bcl-2 positive cells in the PC6, HT7 and GV26 groups, and the myocardial Bcl-2 positive cells in the PC6 and HT7 groups were significantly increased relevant to the model group (P<0.01, P<0.05). No significant changes were found in the KI 6 group in the cell apoptosis index and percentages and Bax- and Bcl-2-positive cells of both myocardium and cerebral cortex tissues compared with the model group (P>0.05)．. CONCLUSION: EA stimulation of PC6 and HT7 can inhibit CI injury induced cell apoptosis of cerebral and myocardial tissues in CI rats, which is possibly associated with its effects in down-regulating Bax expression and up-regulating Bcl-2 expression of both myocardial and cerebral tissues.

MOXIBUSTION ALLEVIATES GASTRIC PRECANCEROUS LESIONS IN RATS BY PROMOTING CELL APOPTOSIS AND INHIBITING PROLIFERATION-RELATED ONCOGENES.

BACKGROUND: It is well known that gastric mucosa dysplasia and intestinal metaplasia are gastric precancerous lesions (GPL). Moxibustion treatment of Liangmen (ST21) and Zusanli (ST36) alleviated the inflammatory response and dysplasia of gastric mucosa in our previous study. The purpose of this study was to further examine the underlying mechanism of moxibustion treatment of ST21 and ST36 on GPL. MATERIALS AND METHODS: Sixty SD rats were divided into five groups and rats with GPL were treated with either moxibustion (ST), moxibustion (Sham), or vitacoenzyme. B-cell lymphoma 2 (bcl-2), tumor protein p53 (P53) and cellular Myc (C-MYC), which are related to cell apoptosis, proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), argyrophilic nucleolar organizer region proteins (Ag-NORs), which are associated with cell proliferation, and cell signaling proteins, nuclear factor kappa B (NF-κB), epidermal growth factor receptor (EGFR) and phosphorylated extracellular signal regulated kinase (p-ERK), were measured after moxibustion treatment. RESULTS: Compared with Control group, gastric mucosa in GPL group showed abnormal mucosal proliferation and pathological mitotic figure, the mRNA expression of bcl-2, P53 and C-MYC increased significantly (P < 0.01), the protein expression of PCNA, VEGF, Ag-NORs and the activity of NF-κß as well as EGFR/ERK signaling proteins also increased significantly (P < 0.01). Moxibustion treatment decreased gastric mucosal proliferation and pathological mitotic figure, down-regulated the mRNA expression of bcl-2, P53, C-MYC (P < 0.01), decreased the protein expression of PCNA, VEGF, Ag-NORs and the activity of NF-κß as well as EGFR/ERK signaling proteins significantly (P < 0.01). But moxibustion treatment of Sham didn't show the same effect on GPL. CONCLUSION: Moxibustion treatment inhibited cell apoptosis and reduced gastric mucosa dysplasia by inhibiting the expression of bcl-2, P53, C-MYC and decreased the activity of NF-κß as well as EGFR/ERK signaling proteins.

NMR-based metabolomics Reveals Alterations of Electro-acupuncture Stimulations on Chronic Atrophic Gastritis Rats.

Chronic atrophic gastritis (CAG) is a common gastrointestinal disease which has been considered as precancerous lesions of gastric carcinoma. Previously, electro-acupuncture stimulation has been shown to be effective in ameliorating symptoms of CAG. However the underlying mechanism of this beneficial treatment is yet to be established. In the present study, an integrated histopathological examination along with molecular biological assay, as well as 1H NMR analysis of multiple biological samples (urine, serum, stomach, cortex and medulla) were employed to systematically assess the pathology of CAG and therapeutic effect of electro-acupuncture stimulation at Sibai (ST 2), Liangmen (ST 21), and Zusanli (ST 36) acupoints located in the stomach meridian using a rat model of CAG. The current results showed that CAG caused comprehensive metabolic alterations including the TCA cycle, glycolysis, membrane metabolism and catabolism, gut microbiota-related metabolism. On the other hand, electro-acupuncture treatment was found able to normalize a number of CAG-induced metabolomics changes by alleviating membrane catabolism, restoring function of neurotransmitter in brain and partially reverse the CAG-induced perturbation in gut microbiota metabolism. These findings provided new insights into the biochemistry of CAG and mechanism of the therapeutic effect of electro-acupuncture stimulations.

[Effect of moxa-burning heat stimulating Liangmen (ST 21) and Zusanli (ST 36) on proliferation and apoptosis signaling proteins in rats with stress-induced gastric ulcer].

OBJECTIVE: To observe the effect of moxa-burning heat stimulating acupoints of Liangmen (ST 21) and Zusanli (ST 36) on the proliferation and apoptosis signaling proteins in rats with stress-induced gastric ulcer. METHODS: Forty rats were randomly divided into four groups: negative control (NC), ulcer control (UC), acupoints of stomach meridian (ASM), and acupoints control (AC). The acute gastric ulcer model was established by bound and water immersion. Rats in NC and UC groups didn't receive any moxa-burning heat stimulating treatment, while rats in ASM and AC groups were treated with buringmoxa heat stimulating the acupoints of Liangmen (ST 21) and Zusanli (ST 36) and their controlled points, respectively. Rats in all groups were sacrificed after 12 consecutive days treatment. The ulcer index was evaluated by using Guth's method. The expression of tumor necrosis factor-alpha (TNF-α), apoptotic protease activating facter-1 (Apaf-1), Caspase-3, p21 activated kinase 1 (PAK1), extracellular regulated protein kinases 2 (ERK2), phosphorylated ERK2 (pERK2), phosphoinositide 3-kinase (PI3K) and RAC-alpha serine/threonine-protein kinase (Akt) in gastric mucosa was detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with UC group, the ulcer index of ASM and AC groups decreased, and the injured gastric mucosa was improved, the expression of TNF-α, Apaf-1 and Caspase-3 in gastric mucosa was significantly reduced (P < 0.05), while the expression of PAK1, ERK2, pERK2, PI3K and Akt in gastric mucosa was significantly increased (P < 0.05). And ASM showed better effect than AC group (P < 0.05). CONCLUSION: Moxa-burning Heat stimulating of Liangmen (ST 21) and Zusanli (ST 36) could promote the recovery of gastric mucosal lesion probably by inhibiting cell apoptosis and promoting cell proliferation in stress-induced gastric ulcer.

[Action mechanism of electroacupuncture at stomach meridian acupoints for oxidative damage in rats with gastric ulcer].

OBJECTIVE: To observe the effects of electroacupuncture (EA) at stomach meridian acupoints on expression of oxidation damage factors in serum and gastric mucosal cells in rats with gastric ulcer, and to explore the mechanism of EA at stomach meridian acupoints for oxidative damage in rats with gastric ulcer. METHODS: Forty clean-grade SD rats were randomly divided into a normal group, a model group, a stomach meridian group and a gallbladder meridian group, ten rats in each one. Except the normal group, rats in the remaining groups were applied the restraint-cold stress method to establish the model of gastric ulcer. Rats in the normal group and model group received no treatment; rats in the stomach meridian group were treated with EA at "Liangmen" (ST 21) and "Zusanli" (ST 36); rats in the gallbladder meridian group were treated with EA at "Riyue" (GB 24) and "Yanglingquan" (GB 34). The EA was given for 30 min, once a day for 7 days totally. The change of gastric mucosal morphology was observed by routine light microscope; enzyme linked immunosorbent assay was used to detect the expressions of malondialdehyde (MDA), glutathione peroxidase (GSH-px) and tumor necrosis factor-α (TNF-α), interleukin-2(IL-2), interleukin-6(IL-6) in serum and gastric mucosal cells of rats. RESULTS: After treatment, compared with the model group, the gastric mucosal damage index was decreased in the stomach meridian group and gallbladder meridian group (both P<0.05), the expressions of MDA, TNF-α, IL-2 and IL-6 in serum and gastric mucosal cells were significantly decreased in the stomach meridian group (all P<0.01), but the contents of GSH-Px in serum and gastric mucosal cells were increased significantly (both P<0.01). Compared with the gallbladder meridian group, the gastric mucosal damage index as well as the expressions of MDA,TNF-α, IL-2 and IL-6 in serum and gastric mucosal cells were significantly decreased in the stomach meridian group rats (P<0.05, P<0.01), and the contents of GSH-px in serum and gastric mucosal cells were increased significantly (both P<0.01). CONCLUSIONS: EA at stomach meridian acupoints is likely to inhibit the expressions of oxidative damage factors to promote the repair of gastric mucosal injury, which indicates the correlation between meridians and zang-fu.

A combined dynamic inhalation device designed for moxa-smoking toxicity testing.

OBJECTIVE: To design a combined dynamic inhalation device for testing the toxicity induced by moxa smoking. METHODS: The new apparatus (Patent No. 201120101911.5) includes air renewal and recycling systems, a gas generating device, a gas control unit, and a device to measure and control temperature and humidity. Sprague-Dawley rats were tested for acute and sub-chronic toxicity after exposure to moxa-burning smoke. RESULTS: We found an LC50 of 1.2 x 10(4) mg/m3 in the acute toxicity assays. In sub-chronic toxicity tests the organ coefficients studied showed no significant differences within rats groups of the same gender after treatment with moxa smoke or a month of recovery. However, mean gray degree of lung 70 heat shock protein (HSP70) was significantly elevated in the high dose group in comparison with the low dose group (P < 0.05); mean gray degree, mean optical density, gross area of HSP70 in other organs and caspase-9 parameters showed no significant differences between groups. CONCLUSION: These results suggest that moxa smoke had no overt toxicity in rats. This work provides evidence and reference for the design of dynamic inhalation exposure systems.

Neuron discharge and c-Fos expression in the nucleus of the solitary tract following electroacupuncture at acupoints of the Yangming Stomach Meridian of Foot.

The nucleus of the solitary tract (nucleus tractus solitarii; NTS) is a primary center for both visceral afferents and somatic afferents. Previous experiments have demonstrated that the NTS is closely connected to the stomach and acupoints in the Yangming Stomach Meridian of Foot (ST Meridian). In this study, extracellular recording and immunochemistry methods were used to analyze the discharge of neurons and c-Fos protein expression in the NTS following acupuncture at different acupoints and a nonacupoint. A total of 104 discharging neurons were detected in the NTS of 52 rats, of which 86 provided complete data. After acupuncture at Sibai (ST 2), Zusanli (ST 36), Neiting (ST 44), Quanliao (SI 18), and the nonacupoint, the neuron response rate in the NTS was 65.12%, 51.16%, 46.51%, 34.88% and 31.40% respectively. For neuron response rate, there was a significant difference among Sibai (ST 2), Zusanli (ST 36), Neiting (ST 44), Quanliao (SI 18), and the nonacupoint (p < 0.01 or p < 0.05). In the other 48 rats, the number of c-Fos immunoreactive neurons in the NTS by electroacupuncture (EA) at Sibai (ST 2) group was significantly higher than that EA at other acupoints and the nonacupoint (p < 0.05 or p < 0.01). EA at both Zusanli (ST 36) and Neiting (ST 44) increased c-Fos immunoreactive neurons significantly over EA at Quanliao (SI 18) and the nonacupoint (p < 0.05 or p < 0.01), while there was no difference between EA at Quanliao (SI 18) and the nonacupoint group (p > 0.05). The experiments demonstrated that the afferent convergence in NTS are different by body surface points stimulus, which suggests that the NTS might be a primary center in the central nervous system receiving acupoints stimulus from the ST Meridian.

[Effects of electroacupuncture of acupoints of pericardium meridian on serum VEGF content and cerebral VEGF expression in cerebral ischemia rats].

OBJECTIVE: To observe the effect of Electroacupuncture (EA) of "Quze" (PC 3) and "Neiguan" (PC 6) on cerebral angiogenesis in cerebral ischemia (CI) rats, so as to research its mechanisms underlying improvement of stroke. METHODS: Fifty SD rats were randomized into normal control, sham, model, EA-PC 3-PC 6, and EA-Hegu (LI 4)-Quchi (LI 11) groups. CI model was established by occlusion of the middle cerebral artery. EA (2-4 V, 20 Hz) was applied to PC3, PC6, LI 4 and LI 11 respectively 6 h, 24 h, 48 h and 72 h after modeling, 30 min for each time. Serum vascular endothelial growth factor (VEGF) content was assayed by enzyme linked immunosorbent assay (ELISA), and cerebral VEGF immunoactivity was detected by immunohistochemistry. RESULTS: Compared to the normal control group, serum VEGF content, cerebral VEGF expression level, and the number of VEGF-positive microvessels in the ischemic cerebral region in the model, EA-PC 3-PC 6 and EA-LI 4-LI 11 groups were increased significantly (P<0.01). In comparison with the model group, serum VEGF content, cerebral VEGF expression level, and the number of VEGF-positive microvessels in the EA-LI 4-LI 11 and EA-PC 3-PC 6 groups were up-regulated further (P<0.01), and the effects of the EA-PC 3-PC 6 group were obviously superior to those of the EA-LI4-LI 11 group in up-regulating VEGF expression and the number of VEGF-positive microvessels (P<0.01). No significant difference was found between the EA-LI 4-LI 11 and EA-PC 3-PC 6 groups in serum VEGF levels (P>0.05). CONCLUSION: EA of acupoints of both Large Intestine Meridian and Pericardium Meridian can up-regulate serum VEGF, cerebral VEGF expression and the number of VEGF-positive microvessels in CI rats, suggesting an increase of cerebral angiogenesis after the treatment. The effect of EA-PC 3-PC 6 is markedly superior to that of EA-LI 4-LI 11.

[Effect of moxibustion on motility, absorption and content of ATP in small intestine of spleen-deficiency rats].

OBJECTIVE: To explore the mechanism of reinforcing function of moxibustion to spleen-stomach. METHODS: Forty healthy Sprague Dawley rats were randomly divided into 4 groups: group A (blank group), group B (model group), group C (moxibustion group) and group D (herbs group). The rat model of spleen-deficiency was established by intragastric administration with 200% Dahuang (Rhubarb) infusion. The rats in group A and B, and D served as the blank control, model, and Sijunzi decoction group respectively, while those in group C received moxibustion at "Zusanli" (ST 36), "Zhongwan" (CV 12), "Guanyuan" (CV 4), "Pishu" (BL 20) and "Weishu" (BL 21), etc. The common symptoms and intestinal propulsive rate were observed. The content of I-xylose in serum was detected by phloroglucinol method. Colorimetry method was used to detected content of ATP in jejunum tissues. RESULTS: Compared with group A, the symptom score in group B was increased significantly (both P < 0.01), while the intestinal propulsive rates, the content of D-xylose in serum and ATP in jejunum tissues were decreased significantly (P < 0.05, P < 0.01). Compared with group B, the symptom score in group C and D was decreased significantly (both P < 0.01), while the intestinal propulsive rates, the content of D-xylose in serum and ATP in jejunum tissues were increased significantly (P < 0.05, P < 0.01). There were no significant difference between group C and D (P > 0.05). CONCLUSION: Moxibustion at "Zusanli" (ST 36) etc. could relieve symptoms of spleen-deficiency, enhance motility and absorption functions of small intestine and improve metabolism of small intestine. The efficacy is equal to administration of Sijunzi decoction.

[Effects of moxibustion on gastric mucosal EGF and TGF-alpha contents and epidermal growth factor receptor expression in rats with gastric mucosal lesion].

OBJECTIVE: To observe the effects of moxibustion on contents of epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) and epidermal growth factor receptor (EGFR) expression in the gastric mucosa tissue in rats with gastric mucosal lesion. METHODS: Thirty-six SD rats were randomly and equally divided into control, model and moxibustion groups. Gastric mucosal lesion model was duplicated by restraint and cool water immersion stress. Pre-moxibustion was applied to "Zusanli" (ST 36) and "Zhongwan" (CV 12), "Pishu" (BL 20) and "Weishu" (BL 21) alternately, once everyday for 8 days before modeling. The contents of EGF and TGF-alpha in gastric mucosa were detected by enzyme-linked immunosorbent assay (ELISA) and the expression of EGFR determined by immunohistochemistry. RESULTS: Compared with the control group, only TGF-alpha content in the gastric mucosa in the model group was increased significantly (P < 0.05). Compared with the model group, the EGF and TGF-alpha contents and EGFR immunoactivity in the gastric mucosa were increased significantly in the moxibustion group (P < 0.05, P < 0.01). CONCLUSION: Pre-moxibustion at ST 36, CV 12, BL 20 and BL 21 can up-regulate gastric mucosal EGF and TGF-alpha contents and EGFR protein expression in gastric mucosa lesion rats, which may contribute to its effect in relieving stress-induced gastric mucosal injury.

[Effect of electroacupuncture of "Neiguan" (PC 6) on Na+-K+-ATPase activity and its gene expression in cardiocyte membrane in rats with myocardial ischemia-reperfusion injury].

OBJECTIVE: To compare the effects of electroacupuncture (EA) of "Neiguan" and other acupoints on Na+ -K+ -ATPase activity and its gene expression in ischemic myocrardial cell membrane in rats. METHODS: A total of 50 Wistar rats were randomly divided into pseudo-operation (control), model, Neiguan (PC6), Shenmen (HT7) and Hegu (LI4) groups, with 10 rats in each group. Myocardial ischemia-reperfusion (I/R) model was duplicated by occlusion of the descending anterior branch of the left coronary artery and reperfusion. EA (30/100 Hz, 2-4 mA) was applied to the above-mentioned acupoints for 20 min respectively. Na+ -K+ -ATPase activity was determined by inorganic phosphorus colorimetry, and Na+ -K+ -ATPase gene expression was detected using reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: Compared with control group, the activity of Na+ -K+ -ATPase in model, PC6, HT7 and LI4 groups decreased significantly after myocardial I/R (P<0.05, 0.01); while in comparison with model, the activity of Na+ -K+ -ATPase in PC6 group increased considerably (P<0.01). Regarding the expression of Na+ -K+ -ATPase mRNA, compared with control group, it was down-regulated significantly in model, HT7 and LI4 groups (P<0.01); while in PC6 group, Na+ -K+ -ATPase mRNA expression was up-regulated markedly compared with model group (P<0.05); The activity of Na+ -K+ -ATPase and the expression of Na+ -K+ -ATPase mRNA in HT7 and LI4 groups were significantly lower than those in PC6 group (P<0.01, 0.05). No significant differences were found between HT7 and LI4 groups in the activity of Na+ -K+ -ATPase and the expression of Na+ -K+ -ATPase mRNA (P>0.05). CONCLUSION: EA of "Neiguan" (PC6) may potentiate the activity of Na+ -K+ -ATPase and up-regulate Na+ -K+ -ATPase mRNA expression, which may contribute to its protective effect on ischemic cardiocytes.

[Expressions of epidermal growth factor receptor signaling substances in gastric mucosal cells influenced by serum derived from rats treated with electroacupuncture at stomach meridian acupoints].

OBJECTIVE: To study the effects of serum derived from rats treated with electroacupuncture at stomach meridian acupoints on the expressions of epidermal growth factor receptor (EGFR) signaling substances phospholipase C gamma-1 (PLC gamma-1), protein kinase C (PKC) and c-myc in gastric mucosal cells. METHODS: Sixty rats were randomly divided into normal group, stomach meridian group, gallbladder meridian group, stomach meridian plus PD153035 group and gallbladder meridian plus PD153035 group. Water-immersion and restrained stress methods were adopted for inducing gastric mucosal injury in the rats. Gastric mucosal cells were separated by using pronase digestion method, and incubated by PD153035, a EGFR inhibitor, and 100 ml/L serum. The expression of PLC gamma-1 in the gastric mucosal cells was tested by enzyme linked-immunosorbent assay (ELISA), while the expression of PKC by isotope incorporate assay and the expression of c-myc by reverse transcription polymerase chain reaction assay (RT-PCR). RESULTS: In gastric mucosal cells, weak expressions of PLC gamma-1, PKC and c-myc were seen in the normal group, and relatively strong expressions of PLC gamma-1, PKC and c-myc were seen in the stomach meridian group and the gallbladder meridian group, among which, the expressions of PLC gamma-1, PKC and c-myc in the stomach meridian group were the strongest, and there was a significant difference between the stomach meridian group and the gallbladder meridian group (P<0.01). Relative weak expressions of PLC gamma-1, PKC and c-myc were seen in the stomach meridian plus PD153035 group and the gallbladder meridian plus PD153035 group, and there was a significant difference between the stomach meridian group and the stomach meridian plus PD153035 group (P<0.01). CONCLUSIONS: The serum derived from the rats treated with electroacupuncture at stomach meridian acupoints can activate the EGFR singling pathway, and this provides an evidence for the theory of "relative particularity between meridians and viscera" in traditional Chinese medicine.

Effect of pre-moxibustion on apoptosis and proliferation of gastric mucosa cells.

AIM: To observe the effects of pre-moxibustion on apoptosis and proliferation of gastric mucosal cell in rats with stress-induced ulcer, and to analyze the relationship between those effects and the expression of heat-shock protein 70 (HSP70). METHODS: Sixty healthy Sprague Dawley rats were randomly assigned into four groups, namely group A, B, C and D. The animal model of stress ulcer was established by water immersion and restraint stress. The rats in group A, B, and D served as the restraint, model, and non-acupoint controls, respectively, while those in group C received moxibustion at Zusanli and Liangmen points. Immunohistochemical methodology was used to detect the expression of HSP70, apoptosis index (AI, multiply 10(-6)/microm(2)) and proliferation index (PCNA-LI, multiply 10(-6)/microm(2)). The mucosal expression of transforming growth factor alpha(TGF-alpha) was detected by radioimmunoassay. RESULTS: Moxibustion at Zusanli and Liangmen points significantly decreased the gastric injury and the apoptosis of gastric mucosal cells, while markedly increased the mucosal expression of TGF-alpha and HSP70 as well as the proliferation of gastric mucosal cells. Compared with group A, ulcer index (UI) (26.8 +/- 9.8 vs 12.0 +/- 5.9, P < 0.01), AI (9.6 +/- 4.2 vs 4.4 +/- 2.6, P < 0.05) and expression of HSP70 (9.6 +/- 4.2 vs 4.4 +/- 2.6, P < 0.05) were significantly increased, but the content of TGF-alpha (104.7 +/- 51.2 pg/mL vs 254.0 +/- 86.9 pg/mL, P < 0.01) and PCNA-LI (6.9 +/- 4.7 vs 14.9 +/- 4.6, P < 0.05) were significantly decreased in group B. However, ulcer index values (UI) and AI were obviously lower in group C compared to groups B and D (14.1 +/- 5.4 vs 26.8 +/- 9.8 and 26.2 +/- 7.7, P < 0.01; 3.0 +/- 1.6 vs 9.6 +/- 4.2 and 8.2 +/- 5.2, P < 0.05, respectively), but content of TGF-alpha (237.0 +/- 72.6 pg/mL vs 104.7 +/- 51.2 pg/mL and 154.1 +/- 61.3 pg/mL, P < 0.01) and expression of HSP70 (0.13 +/- 0.03 vs 0.08 +/- 0.06 and 0.06 +/- 0.04, P < 0.05) were higher in group C. Furthermore, the PCNA-LI was significantly higher in group C than in group B (21.6 +/- 4.1 vs 6.9 +/- 4.7, P < 0.01). CONCLUSION: Moxibustion at Zusanli and Liangmen points has a protective effect on rats gastric mucosa in stress-induced gastric ulcer, which is closely related to its actions in promoting synthesis of TGF-alpha and proliferation of gastric mucosal cells, suppressing gastric mucosal cell apoptosis, and up-regulating HSP70 expression.

The effect of EA on the gastric mucosal histology and ITF mRNA expression in stress-induced rats.

This study focuses on the effect of electroacupuncture (EA) on the gastric mucosal histology and ITF (intestinal trefoil factor) mRNA in stress-related rat, and the relationship between the gastric protective mechanism of EA at acupoints of Stomach Meridian of Foot-Yangming (SMFY) group and Gallbladder Meridian of Foot-Shaoyang (GMFS) group. Forty rats were randomly divided into 4 groups: blank control group (BCG), model control group (MCG), SMFY group (EA at acupoints of SMFY for 7 days before model inducing), and GMFS group (EA at acupoints of GMFS for 7 days before model inducing). All rats (except normal group) were made model by water immersion and restriction (WRS) on day 7, then the gastric mucosal lesion index (GUI) was accessed, ITF mRNA expression of the tissue was detected by reverse- transcriptase-polymerase chain reaction (RT-PCR) method, and the histological change under light microscope was observed. As a result, the GUI value in SMFY/GMFS groups decreased significantly (p < 0.05 or 0.01). The level of ITF mRNA expression in SMFY group was significantly higher than that in MCG (p < 0.01), while that in GMFS group was higher than MCG but there was no statistical difference (p < 0.05). This result may be due to the intrinsic mechanism of EA's gastric mucosal protection by the upregulation of ITF mRNA expression in gastric mucosal tissue, and the expression variance indicated the classical traditional Chinese medicine (TCM) theory "Relative Particularity between SMFY and Stomach.".

Enhanced expression of epidermal growth factor receptor gene in gastric mucosal cells by the serum derived from rats treated with electroacupuncture at stomach meridian acupoints.

AIM: To investigate the effect of serum derived from rats treated with electroacupuncture at stomach meridian acupoints on the expression of epidermal growth factor receptor (EGFR) gene in gastric mucosal cells. METHODS: The stress-induced gastric mucosal injury in rat model was established by water-immersion and restrained stress methods. 52 rats were randomly divided into: normal group (n = 8), model group (n = 8), model serum group (n = 12), stomach serum group (n = 12), and gallbladder serum group (n = 12). The gastric mucosal cells were separated by pronase-EDTA digestion method and incubated with serum. The EGFR gene expression in gastric mucosal cells was detected by reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: Compared with normal group (0.6860 +/- 0.0594), the serum derived from rats of the stomach group (1.2272 +/- 0.0813, P = 0.00 < 0.01) and gallbladder group (0.9640 +/- 0.0387, P = 0.00 < 0.01) had a tendency to enhance the EGFR gene expression in gastric mucosal cells. Such tendency existed in the model group (0.7104 +/- 0.0457) but with no significant difference (P = 0.495 > 0.05) and in model serum group (0.8516 +/- 0.0409) with an extremely obvious difference (P = 0.001 < 0.01). Furthermore, the EGFR gene expression in stomach serum group was significantly higher than that in gallbladder serum group (P = 0.00 < 0.01). CONCLUSION: The present study shows that serum derived from rats treated with electroacupuncture at stomach meridian acupoints can distinctly increase the EGFR gene expression of gastric mucosal cells. Therefore, there is certain meridian specificity in the serum, which could provide a proof for the TCM theory "particular relation between meridian and internal organ".

Effect of electroacupunture on gastric mucosal intestinal trefoil factor gene expression of stress-induced gastric mucosal injury in rats.

AIM: To investigate electroacupuncture(EA) at the acupoints of Stomach Meridian of Foot-Yangming (SMFY), Gallbladder Meridian of Foot-Yangming (SMFY) on gastric mucosal intestinal trefoil factor (ITF) gene expression detection in stress-induced rats with gastric mucosal lesion, and to explore the regulatory mechanism and significance of EA-related gastric mucosal protective effect. METHODS: Forty rats were randomly divided into 4 groups: Blank group, Model group, Model group+EA at acupoints of SMFY group ("SMFY group"), and Model group+EA at acupoints of GMFY group(GMFY group). All rats (except blank group) were made model by water immersion and restraint stress (WRS). Then the gastric mucosa tissue in each rat was taken off after assessment of gastric mucosal lesion index(GUI), and the expression of ITF mRNA of the tissues was detected by reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: Compared with Model group(54.3+/- 1.34), the GUI value in SMFY group (31+/- 2.21) decreased significantly(P< 0.01), so did that in GMFY group (39.8+/- 1.62, P< 0.05), meanwhile GUI value in SMFY group was significantly lower than in GMFY group(P< 0.01). Compared with Model group (0.65+/- 0.01), EA had a tendency to improve the expression of gastric mucosal ITFmRNA gene: such tendency existed in GMFY group (0.66+/- 0.01) but with no significant difference(P>0.05), in SMFY group(0.76+/- 0.01) with an extremely obvious difference (P< 0.01), furthermore the expression in SMFY group was significantly higher than in GMFY group (P< 0.01). CONCLUSION: The gastric mucosal protective effect by EA at the acupoints of SMFY and GMFY was related to the expression variance of ITF, indicating certain meridian specificity exists. It could be one proof for the TCM theory "Relative particularity between SMFY and stomach".

Expression of c-fos in the nucleus of the solitary tract following electroacupuncture at facial acupoints and gastric distension in rats.

Clinical practice has shown that acupuncture at facial acupoints has curative effects on some visceral diseases (especially gastrointestinal diseases). However, the physiological basis has not been clarified yet. In the present study, expression of c-fos in the nucleus of the solitary tract (NTS) of rats following gastric distension and electroacupuncture (EA) at Yangbai (GB14) and Sibai (ST2) as well as Jiache (ST6) acupoints was observed by using immunohistochemistry technique. After EA at the three facial acupoints, c-fos immunoreactive (c-fos-IR) neurons were mainly distributed in the medial (mNTS) and intermediate subnucleus of the NTS, and a few were scatteredly distributed in the dorsalmedial and commissural subnucleus of the NTS. Furthermore, there is difference in the number of c-fos-IR neurons in the mNTS following EA at the three facial acupoints. The number in the EA at ST2 and GB14 group is the highest and the lowest, respectively. Gastric distension induces obviously the expression of c-fos, which is mainly confined in the mNTS. The results suggest that the noxious visceral and somatic afferent information from the stomach and face may converge in the mNTS, which may be involved in the effect of EA at facial acupoints on the gastrointestinal pain.